PROJECT SUMMARY Genome-wide association studies (GWAS) provide insight to underlying etiologies of disease not obvious through clinical evaluation or pathophysiology alone. Genetic variants associated with complex, often refractory diseases such as Alzheimer’s Disease (AD) and Multiple Sclerosis (MS) may hold the key for the next generation of treatment options. Leveraging genetic data with current standards of care and known pathophysiology can provide a strong premise for mechanistic studies and novel drug targets. My current work studies the molecular genetics of CD33 in AD under Dr. Steve Estus. CD33 normally acts to inhibit microglial activation in the brain, suppressing amyloid clearance. We are investigating how the AD-protective single nucleotide polymorphism (SNP) in CD33 modulates protein, and thereby cellular, function. This SNP leads to an increase in an alternative CD33 protein isoform which, based on our recent genetic data, may promote—rather than suppress—microglial activation. I will learn new technical skills during the F99 phase of this award, and I will use these skills as I switch focus to MS and progress into the K00 phase to acquire additional, powerful techniques and models including work with patient samples, pluripotent stem cells, single-cell sequencing technologies, and animal models. In Specific Aim 1, I detail how my training in molecular biology and genetic concepts has allowed me to conceive independent hypotheses and carry out complex experiments. My doctoral work on the molecular genetics of CD33 and its association with reduced AD risk has provided training in GWAS interpretation, quantitative PCR, immunoassays such as Western blotting and co-immunoprecipitation, and genetic techniques including transfection and genome editing strategies in cell culture. In Specific Aim 2, I will continue to develop as a scientist and finish my doctoral work, carrying out increasingly complex studies to include high-resolution confocal imaging and subcellular localization, measuring time- and dose-dependent protein phosphorylation in situ, gene expression arrays, and functional assays including phagocytosis in vitro. I will also continue developing my professional skills such as oral and written communication, networking, and mentorship. In Specific Aim 3, I will extend my doctoral training to include work with human tissue samples and mouse models of MS. I have not yet identified a specific postdoctoral mentor, but my ideal mentor will have experience conducting human subjects research, using mouse models of MS, and have a strong track record of training fellows to become tenure-track faculty. I will identify a mentorship team to guide my technical and professional development during this phase. I will leverage my current training in molecular genetics to identify MS-associated functional SNPs, my training-in-progress to identify the mechanism behind these SNPs at the protein and intracellular signaling levels, and my future ...